Anti-CD146 antibody [P1H12]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD146 antibody [P1H12] (AB24577)

IHC image of CD146 staining in human aorta formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab24577, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

Lab

Western blot - Anti-CD146 antibody [P1H12] (AB24577)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% BSA before being incubated with ab24577 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-CD146 antibody [P1H12] (ab24577) at 5 µg/mL

All lanes:

HUVEC (Human Umbilical Vein Endothelial Cell) whole cell lysate at 25 µg

Secondary

All lanes:

Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

Predicted band size: 72 kDa

Observed band size: 110 kDa

true

Exposure time: 20min

• WB

Lab

Western blot - Anti-CD146 antibody [P1H12] (AB24577)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% BSA before being incubated with ab24577 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-CD146 antibody [P1H12] (ab24577) at 5 µg/mL

All lanes:

Human blood vessel: artery normal tissue lysate - membrane extract (<a href='/en-us/products/unavailable/human-blood-vessel-artery-normal-tissue-lysate-membrane-extract-ab28989'>ab28989</a>) at 25 µg

Secondary

All lanes:

Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

Predicted band size: 72 kDa

Observed band size: 110 kDa

true

Exposure time: 1min